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CD3?/? in sea bass (Dicentrarchus labrax): Molecular characterization and expression analysis.

ABSTRACT: The CD3 complex is the common marker on the surface of both ?? and ?? T cells and is essential for formation of the T-cell receptor complex and for T-cell activation. In this paper, we report the gene cloning and molecular characterization of a CD3?/? homologue in sea bass (Dicentrarchus labrax), the analysis of transcription levels in lymphoid and non-lymphoid organs and the gene regulation after in vitro stimulation with LPS and PHA. Four cysteine residues in the extracellular domain, involved in the constitution of immunoglobulin-like domain, are present in sea bass CD3?/? sequence and they are conserved both in number and position from mammals to teleost sequences. Similar to other known CD3?/?s, in sea bass CD3?/? there is also a conserved immunoreceptor tyrosine-based activation ITAM motif that could be responsible for its individual signal transduction capacity. The real time RT-PCR basal analysis shows the highest level of CD3?/? mRNA in thymus, followed by peripheral blood leucocytes, spleen, gills, gut, liver, head kidney, brain and muscle. The expression analysis under stimuli condition reveals a significant decrease of CD3?/? expression after LPS stimulation and a significant increase after PHA-L stimulation, in agreement with mammals results. In conclusion, these data allow us to affirm that sea bass CD3?/? can be used as a T cell marker and will help in adding new insight on the immune response mechanisms of sea bass.

SUBMITTER: Randelli E

PROVIDER: S-EPMC3787811 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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CD3γ/δ in sea bass (Dicentrarchus labrax): Molecular characterization and expression analysis.

Randelli Elisa E Scapigliati Giuseppe G Buonocore Francesco F

Results in immunology 20110830 1

The CD3 complex is the common marker on the surface of both αβ and γδ T cells and is essential for formation of the T-cell receptor complex and for T-cell activation. In this paper, we report the gene cloning and molecular characterization of a CD3γ/δ homologue in sea bass (Dicentrarchus labrax), the analysis of transcription levels in lymphoid and non-lymphoid organs and the gene regulation after in vitro stimulation with LPS and PHA. Four cysteine residues in the extracellular domain, involved ...[more]

PMID: 24371550

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Project description:BackgroundImmunoglobulins (Igs) are fundamental components of the adaptive immune system of vertebrates, with the IgT/IgZ isotype specific of Teleosts. In this paper we describe the identification of an IgT heavy chain from the European sea bass (Dicentrarchus labrax L.), its molecular characterization and tissue mRNA localization by in situ hybridization.ResultsSea bass IgT consists of 552 aa (Accession Number KM410929) and it contains a putative 19 amino acids long signal peptide and one potential N-glycosylation site. The C-region consists of four CH domains; each contains the cysteine and tryptophan residues required for their correct folding. Based on the recent sequencing of sea bass genome, we have identified five different genomic contigs bearing exons unequivocally pertaining to IgT (CH2, CH3 and CH4), but none corresponded to a complete IgH locus as IgT sequences were found in the highly fragmented assembled genomic regions which could not be assigned to any major scaffold. The 3D structure of sea bass IgT has been modelled using the crystal structure of a mouse Ig gamma as a template, thus showing that the amino acid sequence is suitable for the expected topology referred to an immunoglobulin-like architecture. The basal expression of sea bass IgT and IgM in different organs has been analysed: gut and gills, important mucosal organs, showed high IgT transcripts levels and this was the first indication of the possible involvement of sea bass IgT in mucosal immune responses. Moreover, sea bass IgT expression increased in gills and spleen after infection with nodavirus, highlighting the importance of IgT in sea bass immune responses. In situ hybridization confirmed the presence of IgT transcripts in the gut and it revealed a differential expression along the intestinal tract, with a major expression in the posterior intestine, suggesting the hindgut as a site for the recruitment of IgT+ cells in this species. IgT transcripts were also found in gill filaments and parallel lamellae and, for the first time, we identified scattered IgT positive cells in the liver, with a strong signal in the hepatic parenchyma.ConclusionsIn conclusion, we performed a full molecular characterization of IgT in sea bass that points out its possible involvement in mucosal immune responses of this species.

Dataset Information

CD3?/? in sea bass (Dicentrarchus labrax): Molecular characterization and expression analysis.

Publications

CD3γ/δ in sea bass (Dicentrarchus labrax): Molecular characterization and expression analysis.

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